TY  - JOUR
AU  - Aničić, Neda
AU  - Patelou, Efstathia
AU  - Papanikolaou, Antigoni
AU  - Kanioura, Anthi
AU  - Valdesturli, Camilla
AU  - Arapitsas, Panagiotis
AU  - Skorić, Marijana
AU  - Dragićević, Milan
AU  - Gašić, Uroš
AU  - Koukounaras, Athanasios
AU  - Kostas, Stefanos
AU  - Sarrou, Eirini
AU  - Martens, Stefan
AU  - Mišić, Danijela
AU  - Kanellis, Angelos
PY  - 2021
UR  - www.frontiersin.org
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/4219
AB  - Cistus creticus L. subsp. creticus (rockrose) is a shrub widespread in Greece and the Mediterranean basin and has been used in traditional medicine as herb tea for colds, for healing and digestive hitches, for the treatment of maladies, as perfumes, and for other purposes. Compounds from its flavonoid fraction have recently drawn attention due to antiviral action against influenza virus and HIV. Although several bioactive metabolites belonging to this group have been chemically characterized in the leaves, the genes involved in their biosynthesis in Cistus remain largely unknown. Flavonoid metabolism during C. creticus fruit development was studied by adopting comparative metabolomic and transcriptomic approaches. The present study highlights the fruit of C. creticus subsp. creticus as a rich source of flavonols, flavan-3-ols, and proanthocyanidins, all of which displayed a decreasing trend during fruit development. The majority of proanthocyanidins recorded in Cistus fruit are B-type procyanidins and prodelphinidins, while gallocatechin and catechin are the dominant flavan-3-ols. The expression patterns of biosynthetic genes and transcription factors were analyzed in flowers and throughout three fruit development stages. Flavonoid biosynthetic genes were developmentally regulated, showing a decrease in transcript levels during fruit maturation. A high degree of positive correlations between the content of targeted metabolites and the expression of biosynthetic genes indicated the transcriptional regulation of flavonoid biosynthesis during C. creticus fruit development. This is further supported by the high degree of significant positive correlations between the expression of biosynthetic genes and transcription factors. The results suggest that leucoanthocyanidin reductase predominates the biosynthetic pathway in the control of flavan-3-ol formation, which results in catechin and gallocatechin as two of the major building blocks for Cistus proanthocyanidins. Additionally, there is a decline in ethylene production rates during non-climacteric Cistus fruit maturation, which coincides with the downregulation of the majority of flavonoid- and ethylene-related biosynthetic genes and corresponding transcription factors as well as with the decline in flavonoid content. Finally, functional characterization of a Cistus flavonoid hydroxylase (F3′5′H) was performed for the first time.
PB  - Frontiers Media S.A.
T2  - Frontiers in Plant Science
T1  - Comparative Metabolite and Gene Expression Analyses in Combination With Gene Characterization Revealed the Patterns of Flavonoid Accumulation During Cistus creticus subsp. creticus Fruit Development
VL  - 12
DO  - 10.3389/fpls.2021.619634
SP  - 385
ER  - 

@article{
author = "Aničić, Neda and Patelou, Efstathia and Papanikolaou, Antigoni and Kanioura, Anthi and Valdesturli, Camilla and Arapitsas, Panagiotis and Skorić, Marijana and Dragićević, Milan and Gašić, Uroš and Koukounaras, Athanasios and Kostas, Stefanos and Sarrou, Eirini and Martens, Stefan and Mišić, Danijela and Kanellis, Angelos",
year = "2021",
abstract = "Cistus creticus L. subsp. creticus (rockrose) is a shrub widespread in Greece and the Mediterranean basin and has been used in traditional medicine as herb tea for colds, for healing and digestive hitches, for the treatment of maladies, as perfumes, and for other purposes. Compounds from its flavonoid fraction have recently drawn attention due to antiviral action against influenza virus and HIV. Although several bioactive metabolites belonging to this group have been chemically characterized in the leaves, the genes involved in their biosynthesis in Cistus remain largely unknown. Flavonoid metabolism during C. creticus fruit development was studied by adopting comparative metabolomic and transcriptomic approaches. The present study highlights the fruit of C. creticus subsp. creticus as a rich source of flavonols, flavan-3-ols, and proanthocyanidins, all of which displayed a decreasing trend during fruit development. The majority of proanthocyanidins recorded in Cistus fruit are B-type procyanidins and prodelphinidins, while gallocatechin and catechin are the dominant flavan-3-ols. The expression patterns of biosynthetic genes and transcription factors were analyzed in flowers and throughout three fruit development stages. Flavonoid biosynthetic genes were developmentally regulated, showing a decrease in transcript levels during fruit maturation. A high degree of positive correlations between the content of targeted metabolites and the expression of biosynthetic genes indicated the transcriptional regulation of flavonoid biosynthesis during C. creticus fruit development. This is further supported by the high degree of significant positive correlations between the expression of biosynthetic genes and transcription factors. The results suggest that leucoanthocyanidin reductase predominates the biosynthetic pathway in the control of flavan-3-ol formation, which results in catechin and gallocatechin as two of the major building blocks for Cistus proanthocyanidins. Additionally, there is a decline in ethylene production rates during non-climacteric Cistus fruit maturation, which coincides with the downregulation of the majority of flavonoid- and ethylene-related biosynthetic genes and corresponding transcription factors as well as with the decline in flavonoid content. Finally, functional characterization of a Cistus flavonoid hydroxylase (F3′5′H) was performed for the first time.",
publisher = "Frontiers Media S.A.",
journal = "Frontiers in Plant Science",
title = "Comparative Metabolite and Gene Expression Analyses in Combination With Gene Characterization Revealed the Patterns of Flavonoid Accumulation During Cistus creticus subsp. creticus Fruit Development",
volume = "12",
doi = "10.3389/fpls.2021.619634",
pages = "385"
}

Aničić, N., Patelou, E., Papanikolaou, A., Kanioura, A., Valdesturli, C., Arapitsas, P., Skorić, M., Dragićević, M., Gašić, U., Koukounaras, A., Kostas, S., Sarrou, E., Martens, S., Mišić, D.,& Kanellis, A.. (2021). Comparative Metabolite and Gene Expression Analyses in Combination With Gene Characterization Revealed the Patterns of Flavonoid Accumulation During Cistus creticus subsp. creticus Fruit Development. in Frontiers in Plant Science
Frontiers Media S.A.., 12, 385.
https://doi.org/10.3389/fpls.2021.619634

Aničić N, Patelou E, Papanikolaou A, Kanioura A, Valdesturli C, Arapitsas P, Skorić M, Dragićević M, Gašić U, Koukounaras A, Kostas S, Sarrou E, Martens S, Mišić D, Kanellis A. Comparative Metabolite and Gene Expression Analyses in Combination With Gene Characterization Revealed the Patterns of Flavonoid Accumulation During Cistus creticus subsp. creticus Fruit Development. in Frontiers in Plant Science. 2021;12:385.
doi:10.3389/fpls.2021.619634 .

Aničić, Neda, Patelou, Efstathia, Papanikolaou, Antigoni, Kanioura, Anthi, Valdesturli, Camilla, Arapitsas, Panagiotis, Skorić, Marijana, Dragićević, Milan, Gašić, Uroš, Koukounaras, Athanasios, Kostas, Stefanos, Sarrou, Eirini, Martens, Stefan, Mišić, Danijela, Kanellis, Angelos, "Comparative Metabolite and Gene Expression Analyses in Combination With Gene Characterization Revealed the Patterns of Flavonoid Accumulation During Cistus creticus subsp. creticus Fruit Development" in Frontiers in Plant Science, 12 (2021):385,
https://doi.org/10.3389/fpls.2021.619634 . .

TY  - JOUR
AU  - Trikka, Fotini A.
AU  - Nikolaidis, Alexandros
AU  - Ignea, Codruta
AU  - Tsaballa, Aphrodite
AU  - Tziveleka, Leto-Aikaterini
AU  - Ioannou, Efstathia
AU  - Roussis, Vassilios
AU  - Stea, Eleni A.
AU  - Božić, Dragana
AU  - Argiriou, Anagnostis
AU  - Kanellis, Angelos K.
AU  - Kampranis, Sotirios C.
AU  - Makris, Antonios M.
PY  - 2015
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/2335
AB  - Background: Salvia diterpenes have been found to have health promoting
   properties. Among them, carnosic acid and carnosol, tanshinones and
   sclareol are well known for their cardiovascular, antitumor,
   antiinflammatory and antioxidant activities. However, many of these
   compounds are not available at a constant supply and developing
   biotechnological methods for their production could provide a
   sustainable alternative. The transcriptome of S. pomifera glandular
   trichomes was analysed aiming to identify genes that could be used in
   the engineering of synthetic microbial systems.
   Results: In the present study, a thorough metabolite analysis of S.
   pomifera leaves led to the isolation and structure elucidation of
   carnosic acid-family metabolites including one new natural product.
   These labdane diterpenes seem to be synthesized through miltiradiene and
   ferruginol. Transcriptomic analysis of the glandular trichomes from the
   S. pomifera leaves revealed two genes likely involved in miltiradiene
   synthesis. Their products were identified and the corresponding enzymes
   were characterized as copalyl diphosphate synthase (SpCDS) and
   miltiradiene synthase (SpMilS). In addition, several CYP-encoding
   transcripts were identified providing a valuable resource for the
   identification of the biosynthetic mechanism responsible for the
   production of carnosic acid-family metabolites in S. pomifera.
   Conclusions: Our work has uncovered the key enzymes involved in
   miltiradiene biosynthesis in S. pomifera leaf glandular trichomes. The
   transcriptomic dataset obtained provides a valuable tool for the
   identification of the CYPs involved in the synthesis of carnosic
   acid-family metabolites.
T2  - Bmc Genomics
T1  - Combined metabolome and transcriptome profiling provides new insights
 into diterpene biosynthesis in S. pomifera glandular trichomes
IS  - 935
VL  - 16
DO  - 10.1186/s12864-015-2147-3
ER  - 

@article{
author = "Trikka, Fotini A. and Nikolaidis, Alexandros and Ignea, Codruta and Tsaballa, Aphrodite and Tziveleka, Leto-Aikaterini and Ioannou, Efstathia and Roussis, Vassilios and Stea, Eleni A. and Božić, Dragana and Argiriou, Anagnostis and Kanellis, Angelos K. and Kampranis, Sotirios C. and Makris, Antonios M.",
year = "2015",
abstract = "Background: Salvia diterpenes have been found to have health promoting
   properties. Among them, carnosic acid and carnosol, tanshinones and
   sclareol are well known for their cardiovascular, antitumor,
   antiinflammatory and antioxidant activities. However, many of these
   compounds are not available at a constant supply and developing
   biotechnological methods for their production could provide a
   sustainable alternative. The transcriptome of S. pomifera glandular
   trichomes was analysed aiming to identify genes that could be used in
   the engineering of synthetic microbial systems.
   Results: In the present study, a thorough metabolite analysis of S.
   pomifera leaves led to the isolation and structure elucidation of
   carnosic acid-family metabolites including one new natural product.
   These labdane diterpenes seem to be synthesized through miltiradiene and
   ferruginol. Transcriptomic analysis of the glandular trichomes from the
   S. pomifera leaves revealed two genes likely involved in miltiradiene
   synthesis. Their products were identified and the corresponding enzymes
   were characterized as copalyl diphosphate synthase (SpCDS) and
   miltiradiene synthase (SpMilS). In addition, several CYP-encoding
   transcripts were identified providing a valuable resource for the
   identification of the biosynthetic mechanism responsible for the
   production of carnosic acid-family metabolites in S. pomifera.
   Conclusions: Our work has uncovered the key enzymes involved in
   miltiradiene biosynthesis in S. pomifera leaf glandular trichomes. The
   transcriptomic dataset obtained provides a valuable tool for the
   identification of the CYPs involved in the synthesis of carnosic
   acid-family metabolites.",
journal = "Bmc Genomics",
title = "Combined metabolome and transcriptome profiling provides new insights
 into diterpene biosynthesis in S. pomifera glandular trichomes",
number = "935",
volume = "16",
doi = "10.1186/s12864-015-2147-3"
}

Trikka, F. A., Nikolaidis, A., Ignea, C., Tsaballa, A., Tziveleka, L., Ioannou, E., Roussis, V., Stea, E. A., Božić, D., Argiriou, A., Kanellis, A. K., Kampranis, S. C.,& Makris, A. M.. (2015). Combined metabolome and transcriptome profiling provides new insights
 into diterpene biosynthesis in S. pomifera glandular trichomes. in Bmc Genomics, 16(935).
https://doi.org/10.1186/s12864-015-2147-3

Trikka FA, Nikolaidis A, Ignea C, Tsaballa A, Tziveleka L, Ioannou E, Roussis V, Stea EA, Božić D, Argiriou A, Kanellis AK, Kampranis SC, Makris AM. Combined metabolome and transcriptome profiling provides new insights
 into diterpene biosynthesis in S. pomifera glandular trichomes. in Bmc Genomics. 2015;16(935).
doi:10.1186/s12864-015-2147-3 .

Trikka, Fotini A., Nikolaidis, Alexandros, Ignea, Codruta, Tsaballa, Aphrodite, Tziveleka, Leto-Aikaterini, Ioannou, Efstathia, Roussis, Vassilios, Stea, Eleni A., Božić, Dragana, Argiriou, Anagnostis, Kanellis, Angelos K., Kampranis, Sotirios C., Makris, Antonios M., "Combined metabolome and transcriptome profiling provides new insights
 into diterpene biosynthesis in S. pomifera glandular trichomes" in Bmc Genomics, 16, no. 935 (2015),
https://doi.org/10.1186/s12864-015-2147-3 . .

TY  - JOUR
AU  - Brückner, Kathleen
AU  - Matekalo, Dragana
AU  - Manzano, David
AU  - Papaefthimiou, Dimitra
AU  - Pateraki, Irini
AU  - Scheler, Ulschan
AU  - Ferrer, Albert
AU  - de Vos, Ric C H
AU  - Kanellis, Angelos
AU  - Tissier, Alain
PY  - 2014
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/4287
AB  - Rosemary (Rosmarinus officinalis) produces the phenolic diterpenes carnosic acid and carnosol, which, in addition to their general antioxidant activities, have recently been suggested as potential ingredients for the prevention and treatment of neurodegenerative diseases. Little is known about the biosynthesis of these diterpenes. Here we show that the biosynthesis of phenolic diterpenes in rosemary predominantly takes place in the glandular trichomes of young leaves, and used this feature to identify the first committed steps. Thus, a copalyl diphosphate synthase (RoCPS1) and two kaurene synthase-like (RoKSL1 and RoKSL2) encoding genes were identified and characterized. Expression in yeast (Saccharomyces cerevisiae) and Nicotiana benthamiana demonstrate that RoCPS1 converts geranylgeranyl diphosphate (GGDP) to copalyl diphosphate (CDP) of normal stereochemistry and that both RoKSL1 and RoKSL2 use normal CDP to produce an abietane diterpene. Comparison to the already characterized diterpene synthase from Salvia miltiorrhiza (SmKSL) demonstrates that the product of RoKSL1 and RoKSL2 is miltiradiene. Expression analysis supports a major contributing role for RoKSL2. Like SmKSL and the sclareol synthase from Salvia sclarea, RoKSL1/2 are diterpene synthases of the TPS-e group which have lost the internal gamma-domain. Furthermore, phylogenetic analysis indicates that RoKSL1 and RoKSL2 belong to a distinct group of KSL enzymes involved in specialized metabolism which most likely emerged before the dicot-monocot split.
PB  - Elsevier
T2  - Phytochemistry
T1  - Characterization of two genes for the biosynthesis of abietane-type diterpenes in rosemary (Rosmarinus officinalis) glandular trichomes
VL  - 101
DO  - 10.1016/j.phytochem.2014.01.021
SP  - 52
EP  - 64
ER  - 

@article{
author = "Brückner, Kathleen and Matekalo, Dragana and Manzano, David and Papaefthimiou, Dimitra and Pateraki, Irini and Scheler, Ulschan and Ferrer, Albert and de Vos, Ric C H and Kanellis, Angelos and Tissier, Alain",
year = "2014",
abstract = "Rosemary (Rosmarinus officinalis) produces the phenolic diterpenes carnosic acid and carnosol, which, in addition to their general antioxidant activities, have recently been suggested as potential ingredients for the prevention and treatment of neurodegenerative diseases. Little is known about the biosynthesis of these diterpenes. Here we show that the biosynthesis of phenolic diterpenes in rosemary predominantly takes place in the glandular trichomes of young leaves, and used this feature to identify the first committed steps. Thus, a copalyl diphosphate synthase (RoCPS1) and two kaurene synthase-like (RoKSL1 and RoKSL2) encoding genes were identified and characterized. Expression in yeast (Saccharomyces cerevisiae) and Nicotiana benthamiana demonstrate that RoCPS1 converts geranylgeranyl diphosphate (GGDP) to copalyl diphosphate (CDP) of normal stereochemistry and that both RoKSL1 and RoKSL2 use normal CDP to produce an abietane diterpene. Comparison to the already characterized diterpene synthase from Salvia miltiorrhiza (SmKSL) demonstrates that the product of RoKSL1 and RoKSL2 is miltiradiene. Expression analysis supports a major contributing role for RoKSL2. Like SmKSL and the sclareol synthase from Salvia sclarea, RoKSL1/2 are diterpene synthases of the TPS-e group which have lost the internal gamma-domain. Furthermore, phylogenetic analysis indicates that RoKSL1 and RoKSL2 belong to a distinct group of KSL enzymes involved in specialized metabolism which most likely emerged before the dicot-monocot split.",
publisher = "Elsevier",
journal = "Phytochemistry",
title = "Characterization of two genes for the biosynthesis of abietane-type diterpenes in rosemary (Rosmarinus officinalis) glandular trichomes",
volume = "101",
doi = "10.1016/j.phytochem.2014.01.021",
pages = "52-64"
}

Brückner, K., Matekalo, D., Manzano, D., Papaefthimiou, D., Pateraki, I., Scheler, U., Ferrer, A., de Vos, R. C. H., Kanellis, A.,& Tissier, A.. (2014). Characterization of two genes for the biosynthesis of abietane-type diterpenes in rosemary (Rosmarinus officinalis) glandular trichomes. in Phytochemistry
Elsevier., 101, 52-64.
https://doi.org/10.1016/j.phytochem.2014.01.021

Brückner K, Matekalo D, Manzano D, Papaefthimiou D, Pateraki I, Scheler U, Ferrer A, de Vos RCH, Kanellis A, Tissier A. Characterization of two genes for the biosynthesis of abietane-type diterpenes in rosemary (Rosmarinus officinalis) glandular trichomes. in Phytochemistry. 2014;101:52-64.
doi:10.1016/j.phytochem.2014.01.021 .

Brückner, Kathleen, Matekalo, Dragana, Manzano, David, Papaefthimiou, Dimitra, Pateraki, Irini, Scheler, Ulschan, Ferrer, Albert, de Vos, Ric C H, Kanellis, Angelos, Tissier, Alain, "Characterization of two genes for the biosynthesis of abietane-type diterpenes in rosemary (Rosmarinus officinalis) glandular trichomes" in Phytochemistry, 101 (2014):52-64,
https://doi.org/10.1016/j.phytochem.2014.01.021 . .

TY  - CONF
AU  - Skorić, Marijana
AU  - Todorović, Slađana
AU  - Soković, Marina
AU  - Glamočlija, Jasmina
AU  - Ristić, Mihailo
AU  - Živković, Suzana
AU  - Kanellis, Angelos K
PY  - 2013
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6801
AB  - Cistus creticus subsp. creticus is a native plant of the Mediterranean region and it has been used since an­cient times for its medicinal properties. Predominant compounds in leaves and stems of the C. creticus subsp. creticus, as well as in resin, appear to be labdane type diterpenes, while other terpenes, polyphenols and fla­vonoids contribute to chemical profile of this species. We have implemented in vitro culture of C. creticus, in order to obtain applicable production of secondary metabolites. Successfully established in vitro shoot and root cultures, as well as cultures of transformed roots ("hairy" root cultures) were used for analyses. GC/FID and GC/MS detections were performed to phytochemically characterize the ethanol extracts of shoots, non­transformed and transformed roots from in vitro culture. Labdane diterpenes were the most abundant com­pounds in shoot extracts, but absent from the root extracts. Total phenolic and flavonoid contents were al­so determined in the ethanol extracts of C. creticus shoots, non-transformed and transformed roots. The an­tibacterial assay was carried out using microdilution method in order to determine the minimum inhibitory and bactericidal concentrations (MICs and MBCs) against eight human pathogenic bacteria. For the antifungal bioassays, seven fungi were used. Minimum inhibitory concentration (MIC) and the fungicidal concen­trations (MFCs) were determined using a modified microdilution technique. All tested shoot extract showed better antibacterial activity than streptomycin, and much better antibacterial activity than ampicillin. All root samples (transformed and non-transformed) also showed stronger antibacterial activity than antibiotics with few exceptions. Almost all tested extracts exhibited slightly higher or similar antifungal potential as bifona­zole, and showed much better antifungal effect than ketoconazole.
PB  - Belgrade: Serbian Plant Physiology Society
C3  - Programme and Abstracts: 1st International Conference on Plant Biology and 20th Symposium of the Serbian Plant Physiology Society; 2013 Jun 4-7; Subotica, Belgrade
T1  - Antimicrobial activity of ethanol extracts of in vitro grown Cistus creticus subsp. creticus
SP  - 107
EP  - 108
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6801
ER  - 

@conference{
author = "Skorić, Marijana and Todorović, Slađana and Soković, Marina and Glamočlija, Jasmina and Ristić, Mihailo and Živković, Suzana and Kanellis, Angelos K",
year = "2013",
abstract = "Cistus creticus subsp. creticus is a native plant of the Mediterranean region and it has been used since an­cient times for its medicinal properties. Predominant compounds in leaves and stems of the C. creticus subsp. creticus, as well as in resin, appear to be labdane type diterpenes, while other terpenes, polyphenols and fla­vonoids contribute to chemical profile of this species. We have implemented in vitro culture of C. creticus, in order to obtain applicable production of secondary metabolites. Successfully established in vitro shoot and root cultures, as well as cultures of transformed roots ("hairy" root cultures) were used for analyses. GC/FID and GC/MS detections were performed to phytochemically characterize the ethanol extracts of shoots, non­transformed and transformed roots from in vitro culture. Labdane diterpenes were the most abundant com­pounds in shoot extracts, but absent from the root extracts. Total phenolic and flavonoid contents were al­so determined in the ethanol extracts of C. creticus shoots, non-transformed and transformed roots. The an­tibacterial assay was carried out using microdilution method in order to determine the minimum inhibitory and bactericidal concentrations (MICs and MBCs) against eight human pathogenic bacteria. For the antifungal bioassays, seven fungi were used. Minimum inhibitory concentration (MIC) and the fungicidal concen­trations (MFCs) were determined using a modified microdilution technique. All tested shoot extract showed better antibacterial activity than streptomycin, and much better antibacterial activity than ampicillin. All root samples (transformed and non-transformed) also showed stronger antibacterial activity than antibiotics with few exceptions. Almost all tested extracts exhibited slightly higher or similar antifungal potential as bifona­zole, and showed much better antifungal effect than ketoconazole.",
publisher = "Belgrade: Serbian Plant Physiology Society",
journal = "Programme and Abstracts: 1st International Conference on Plant Biology and 20th Symposium of the Serbian Plant Physiology Society; 2013 Jun 4-7; Subotica, Belgrade",
title = "Antimicrobial activity of ethanol extracts of in vitro grown Cistus creticus subsp. creticus",
pages = "107-108",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6801"
}

Skorić, M., Todorović, S., Soković, M., Glamočlija, J., Ristić, M., Živković, S.,& Kanellis, A. K.. (2013). Antimicrobial activity of ethanol extracts of in vitro grown Cistus creticus subsp. creticus. in Programme and Abstracts: 1st International Conference on Plant Biology and 20th Symposium of the Serbian Plant Physiology Society; 2013 Jun 4-7; Subotica, Belgrade
Belgrade: Serbian Plant Physiology Society., 107-108.
https://hdl.handle.net/21.15107/rcub_ibiss_6801

Skorić M, Todorović S, Soković M, Glamočlija J, Ristić M, Živković S, Kanellis AK. Antimicrobial activity of ethanol extracts of in vitro grown Cistus creticus subsp. creticus. in Programme and Abstracts: 1st International Conference on Plant Biology and 20th Symposium of the Serbian Plant Physiology Society; 2013 Jun 4-7; Subotica, Belgrade. 2013;:107-108.
https://hdl.handle.net/21.15107/rcub_ibiss_6801 .

Skorić, Marijana, Todorović, Slađana, Soković, Marina, Glamočlija, Jasmina, Ristić, Mihailo, Živković, Suzana, Kanellis, Angelos K, "Antimicrobial activity of ethanol extracts of in vitro grown Cistus creticus subsp. creticus" in Programme and Abstracts: 1st International Conference on Plant Biology and 20th Symposium of the Serbian Plant Physiology Society; 2013 Jun 4-7; Subotica, Belgrade (2013):107-108,
https://hdl.handle.net/21.15107/rcub_ibiss_6801 .

TY  - CONF
AU  - Skorić, Marijana
AU  - Todorović, Slađana
AU  - Ristić, Mihailo
AU  - Soković, Marina
AU  - Glamočlija, Jasmina
AU  - Živković, Suzana
AU  - Stojić, Andreja
AU  - Puač, Nevena
AU  - Kanellis, Angelos K
PY  - 2013
UR  - http://terpnet2013.pharm.auth.gr/site/TERPNET_BOOK_OF_ABSTRACTS.pdf
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6320
AB  - Cistus creticus subsp. creticus is a native plant of the Mediterranean region and it has been used since ancient times for its medicinal properties. Labdane type diterpenes are predominant compounds in leaves and stems of the C. creticus subsp. creticus, as well as in resin, while other terpenes, polyphenols and flavonoids contribute to chemical profile of this species. In vitro plant tissue culture is an attractive alternative approach to the traditional methods of plantations, as it offers a controlled supply of biochemicals independent of plant availability and more consistent product quality. In order to obtain applicable production of secondary metabolites of interest we have implemented in vitro culture of several clones of C. creticus, which were previously morphogenetically described. The ethanol extracts of obtained in vitro cultures were analyzed by GC-FID and GC-MS, and were characterized concerning the total phenolic and flavonoid contents. Labdane diterpenes were the most abundant compounds in above ground parts extracts, but absent from the root extracts. Head-space GC-MS was used to determine the profile of volatile compounds in the atmosphere of the culture vessels. α pinene, β pinene, and camphene were the most abundant ones. PTR-MS was used for further quantification of representative volatile compounds in the atmosphere of the culture vessels. Finally, the biological activity of obtained ethanol extract was studied. Microdilution method was used for investigation of antibacterial and antifungal activity. Almost all tested extract against human pathogenic bacteria showed better antibacterial activity than streptomycin, and much better antibacterial activity than ampicillin. All tested extracts also exhibited slightly higher or similar antifungal potential as bifonazole, and showed much better antifungal effect than ketoconazole. Antioxidant activity was determined by using DPPH (2,2-Diphenyl-1-picrylhydrazyl) test, while cytotoxic activity was previously demonstrated by sulforhodamine B (SRB) assay.
PB  - Thessaloniki: Department of Pharmaceutical Science, Aristotle University of Thessaloniki
C3  - TERPNET 2013: 11th International meeting: Biosynthesis, Function and Biotechnology of Isoprenoids in Terrestrial and Marine Organisms; 2013 Jun 1-5; Kolymvari, Greece
T1  - In vitro culture of Cistus creticus subsp. creticus – a source of biological active compounds
SP  - 132
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6320
ER  - 

@conference{
author = "Skorić, Marijana and Todorović, Slađana and Ristić, Mihailo and Soković, Marina and Glamočlija, Jasmina and Živković, Suzana and Stojić, Andreja and Puač, Nevena and Kanellis, Angelos K",
year = "2013",
abstract = "Cistus creticus subsp. creticus is a native plant of the Mediterranean region and it has been used since ancient times for its medicinal properties. Labdane type diterpenes are predominant compounds in leaves and stems of the C. creticus subsp. creticus, as well as in resin, while other terpenes, polyphenols and flavonoids contribute to chemical profile of this species. In vitro plant tissue culture is an attractive alternative approach to the traditional methods of plantations, as it offers a controlled supply of biochemicals independent of plant availability and more consistent product quality. In order to obtain applicable production of secondary metabolites of interest we have implemented in vitro culture of several clones of C. creticus, which were previously morphogenetically described. The ethanol extracts of obtained in vitro cultures were analyzed by GC-FID and GC-MS, and were characterized concerning the total phenolic and flavonoid contents. Labdane diterpenes were the most abundant compounds in above ground parts extracts, but absent from the root extracts. Head-space GC-MS was used to determine the profile of volatile compounds in the atmosphere of the culture vessels. α pinene, β pinene, and camphene were the most abundant ones. PTR-MS was used for further quantification of representative volatile compounds in the atmosphere of the culture vessels. Finally, the biological activity of obtained ethanol extract was studied. Microdilution method was used for investigation of antibacterial and antifungal activity. Almost all tested extract against human pathogenic bacteria showed better antibacterial activity than streptomycin, and much better antibacterial activity than ampicillin. All tested extracts also exhibited slightly higher or similar antifungal potential as bifonazole, and showed much better antifungal effect than ketoconazole. Antioxidant activity was determined by using DPPH (2,2-Diphenyl-1-picrylhydrazyl) test, while cytotoxic activity was previously demonstrated by sulforhodamine B (SRB) assay.",
publisher = "Thessaloniki: Department of Pharmaceutical Science, Aristotle University of Thessaloniki",
journal = "TERPNET 2013: 11th International meeting: Biosynthesis, Function and Biotechnology of Isoprenoids in Terrestrial and Marine Organisms; 2013 Jun 1-5; Kolymvari, Greece",
title = "In vitro culture of Cistus creticus subsp. creticus – a source of biological active compounds",
pages = "132",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6320"
}

Skorić, M., Todorović, S., Ristić, M., Soković, M., Glamočlija, J., Živković, S., Stojić, A., Puač, N.,& Kanellis, A. K.. (2013). In vitro culture of Cistus creticus subsp. creticus – a source of biological active compounds. in TERPNET 2013: 11th International meeting: Biosynthesis, Function and Biotechnology of Isoprenoids in Terrestrial and Marine Organisms; 2013 Jun 1-5; Kolymvari, Greece
Thessaloniki: Department of Pharmaceutical Science, Aristotle University of Thessaloniki., 132.
https://hdl.handle.net/21.15107/rcub_ibiss_6320

Skorić M, Todorović S, Ristić M, Soković M, Glamočlija J, Živković S, Stojić A, Puač N, Kanellis AK. In vitro culture of Cistus creticus subsp. creticus – a source of biological active compounds. in TERPNET 2013: 11th International meeting: Biosynthesis, Function and Biotechnology of Isoprenoids in Terrestrial and Marine Organisms; 2013 Jun 1-5; Kolymvari, Greece. 2013;:132.
https://hdl.handle.net/21.15107/rcub_ibiss_6320 .

Skorić, Marijana, Todorović, Slađana, Ristić, Mihailo, Soković, Marina, Glamočlija, Jasmina, Živković, Suzana, Stojić, Andreja, Puač, Nevena, Kanellis, Angelos K, "In vitro culture of Cistus creticus subsp. creticus – a source of biological active compounds" in TERPNET 2013: 11th International meeting: Biosynthesis, Function and Biotechnology of Isoprenoids in Terrestrial and Marine Organisms; 2013 Jun 1-5; Kolymvari, Greece (2013):132,
https://hdl.handle.net/21.15107/rcub_ibiss_6320 .

TY  - CONF
AU  - Brückner, Kathleen
AU  - Matekalo, Dragana
AU  - Manzano, David
AU  - Papaefthimiou, Dimitra
AU  - Pateraki, Irini
AU  - Boronat, Albert
AU  - Datema, Erwin
AU  - de Vos, Ric C H
AU  - Ferrer, Albert
AU  - Kanellis, Angelos
AU  - Tissier, Alain
PY  - 2013
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/4288
AB  - Plant secondary metabolites contribute substantially to the human pharmacopeae. To better exploit the medicinal plants that produce these compounds, it is necessary to understand how these compounds are synthesized within the plant. Identification of the biosynthetic pathway genes will allow both the breeding of plant varieties with improved productivity and/or metabolic profile, and the development of alternative production methods using biotechnology.
The phenolic diterpenes (PDs) carnosic acid (CA) and carnosol have high antioxidant activities and display potential for the treatment of neurodegenerative disorders. CA and carnosol are found in Rosmarinus and Salvia sp. which are known for their health promoting properties since ages.
As part of the EU-financed TERPMED project, PD biosynthesis is being investigated. To elucidate the pathway, the localization of the biosynthesis of PDs was determined. Glandular trichomes appear to contribute significantly to the biosynthesis of PDs and the transcriptome of glandular trichomes was thus determined by next generation sequencing of cDNA.
A set of gene candidates was selected according to alignment searches and analysis of trichome-specific gene expression profile. The search for terpene synthases yielded two types of candidates for the first steps of the pathway: a copalyl diphosphate synthase (CPS) and kaurene synthase-like (KSL) encoding genes. Genes encoding oxidases of cytochrome P450 clade were the most promising candidates for the downstream steps of the PDs pathway.
Finally, the first steps of the biosynthesis of PDs are reconstituted and further investigations on the role of P450 candidates in downstream steps are being carried out using N. benthamiana transient expression and heterologous expression in yeast.
PB  - Thessaloniki: Department of Pharmaceutical Science, Aristotle University of Thessaloniki
C3  - 11th International meeting Biosynthesis, Function and Biotechnology of Isoprenoids in Terrestrial and Marine Organisms, Book of Abstracts
T1  - Investigating phenolic diterpenes in rosemary (Rosmarinus officinalis) and sage (Salvia fruticosa)
SP  - 59
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_4288
ER  - 

@conference{
author = "Brückner, Kathleen and Matekalo, Dragana and Manzano, David and Papaefthimiou, Dimitra and Pateraki, Irini and Boronat, Albert and Datema, Erwin and de Vos, Ric C H and Ferrer, Albert and Kanellis, Angelos and Tissier, Alain",
year = "2013",
abstract = "Plant secondary metabolites contribute substantially to the human pharmacopeae. To better exploit the medicinal plants that produce these compounds, it is necessary to understand how these compounds are synthesized within the plant. Identification of the biosynthetic pathway genes will allow both the breeding of plant varieties with improved productivity and/or metabolic profile, and the development of alternative production methods using biotechnology.
The phenolic diterpenes (PDs) carnosic acid (CA) and carnosol have high antioxidant activities and display potential for the treatment of neurodegenerative disorders. CA and carnosol are found in Rosmarinus and Salvia sp. which are known for their health promoting properties since ages.
As part of the EU-financed TERPMED project, PD biosynthesis is being investigated. To elucidate the pathway, the localization of the biosynthesis of PDs was determined. Glandular trichomes appear to contribute significantly to the biosynthesis of PDs and the transcriptome of glandular trichomes was thus determined by next generation sequencing of cDNA.
A set of gene candidates was selected according to alignment searches and analysis of trichome-specific gene expression profile. The search for terpene synthases yielded two types of candidates for the first steps of the pathway: a copalyl diphosphate synthase (CPS) and kaurene synthase-like (KSL) encoding genes. Genes encoding oxidases of cytochrome P450 clade were the most promising candidates for the downstream steps of the PDs pathway.
Finally, the first steps of the biosynthesis of PDs are reconstituted and further investigations on the role of P450 candidates in downstream steps are being carried out using N. benthamiana transient expression and heterologous expression in yeast.",
publisher = "Thessaloniki: Department of Pharmaceutical Science, Aristotle University of Thessaloniki",
journal = "11th International meeting Biosynthesis, Function and Biotechnology of Isoprenoids in Terrestrial and Marine Organisms, Book of Abstracts",
title = "Investigating phenolic diterpenes in rosemary (Rosmarinus officinalis) and sage (Salvia fruticosa)",
pages = "59",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_4288"
}

Brückner, K., Matekalo, D., Manzano, D., Papaefthimiou, D., Pateraki, I., Boronat, A., Datema, E., de Vos, R. C. H., Ferrer, A., Kanellis, A.,& Tissier, A.. (2013). Investigating phenolic diterpenes in rosemary (Rosmarinus officinalis) and sage (Salvia fruticosa). in 11th International meeting Biosynthesis, Function and Biotechnology of Isoprenoids in Terrestrial and Marine Organisms, Book of Abstracts
Thessaloniki: Department of Pharmaceutical Science, Aristotle University of Thessaloniki., 59.
https://hdl.handle.net/21.15107/rcub_ibiss_4288

Brückner K, Matekalo D, Manzano D, Papaefthimiou D, Pateraki I, Boronat A, Datema E, de Vos RCH, Ferrer A, Kanellis A, Tissier A. Investigating phenolic diterpenes in rosemary (Rosmarinus officinalis) and sage (Salvia fruticosa). in 11th International meeting Biosynthesis, Function and Biotechnology of Isoprenoids in Terrestrial and Marine Organisms, Book of Abstracts. 2013;:59.
https://hdl.handle.net/21.15107/rcub_ibiss_4288 .

Brückner, Kathleen, Matekalo, Dragana, Manzano, David, Papaefthimiou, Dimitra, Pateraki, Irini, Boronat, Albert, Datema, Erwin, de Vos, Ric C H, Ferrer, Albert, Kanellis, Angelos, Tissier, Alain, "Investigating phenolic diterpenes in rosemary (Rosmarinus officinalis) and sage (Salvia fruticosa)" in 11th International meeting Biosynthesis, Function and Biotechnology of Isoprenoids in Terrestrial and Marine Organisms, Book of Abstracts (2013):59,
https://hdl.handle.net/21.15107/rcub_ibiss_4288 .